16S RNA Gene Research Articles

Effects of polyethylene microplastics on the microbiome and metabolism in larval zebrafish

Various microplastics (MPs) are found in the environment and organisms. MP residues in organisms can affect health; however, their impacts on metabolism in aquatic organisms remain unclear. In this study, zebrafish embryos were exposed to polyethylene MPs with sizes ranging from 1 to 4 μm at concentrations of 0, 10, 100, and 1000 μg/L for 7 days. Through qPCR technology, the results indicated that zebrafish exposed to polyethylene MPs exhibited significant change in microbes of the phyla Firmicutes, Bacteroidetes, Proteobacteria, and Verrucomicrobia, etc. Moreover, 16S RNA gene sequencing revealed that there was a significant difference in alpha diversity between the control and 1000 μg/L MP-treated groups. At the genus level, the abundance of Aeromonas, Shewanella, Microbacterium, Nevskia and Methyloversatilis have increased remarkably. Conversely, the abundance of Pseudomonas, Ralstonia and Stenotrophomonas were significant reduction after MPs exposure. In addition, the levels of TG (triglyceride), TCHO (total cholesterol), NEFA (nonesterified fatty acid), TBA (total bile acid), GLU (glucose) and pyruvic acid significantly changed in MP-treated larval zebrafish, indicating that their metabolism was disturbed by MPs. Transcriptional levels of glucose and lipid metabolism-related genes showed a decreasing trend. Furthermore, LC/MS-based nontargeted metabolomics analysis demonstrated that a total of 59 phospholipid-related substances exhibited significant changes in larval fish treated with 1000 μg/L MPs. The mRNA levels of phospholipid metabolism-related genes were also obviously changed. Pearson correlation analysis indicated that the abundance of Aeromonas, Shewanella and Chitinibacter bacteria showed a negative correlation with most phospholipids, while Nevskia, Parvibacter and Lysobacter showed a positive correlation with most phospholipids. Based on these results, it is suggested that 1–4 μm PE-MPs could impact the microbiome and metabolism of larval zebrafish. All of these results indicated that the health risk of MPs cannot be ignored.

Halotolerant Endophytic Bacterium Serratia rubidaea ED1 Enhances Phosphate Solubilization and Promotes Seed Germination

Quinoa is renowned for its nutritional value and ability to withstand harsh environmental conditions such as salinity. In the present work, we isolated 34 phosphate solubilizing endophytic bacteria associated with the roots of quinoa plants. Based on phosphate solubilization efficiency and biochemical characterization, we selected one isolate named ED1. Ribotyping using partial 16S RNA gene analysis revealed that the selected isolate shares 99.7% identity with Serratia rubidaea. Plant growth promoting (PGP) studies showed that the ED1 strain solubilized complexed forms of phosphate (Ca3(PO4)2). Zinc release from ZnO, Zn3(PO4)2, or ZnCO3 revealed the efficient ZnO solubilization by the ED1 strain. Except for proteases, the strain ED1 produced siderophores, cellulase, ammonia and exhibited oligonitrophilic features. Indole acetic acid (IAA) production was detected with and without the L-tryptophan precursor. Next, we demonstrated that the ED1 strain tolerated 1.5 M NaCl final concentration and exhibited intrinsic resistance to seven antibiotics frequently prescribed for medical use. Moreover, we found that ED1 strain withstood 2 mg/L of Cadmium and 1 mg/L of either Nickel or Copper. Furthermore, we observed that S. rubidaea ED1 stimulated quinoa seeds germination and seedlings growth under salt stress conditions. Lastly, we discuss the advantages versus disadvantages of applying the S. rubidaea ED1 strain as a beneficial agent for salty and/or heavy metals contaminated soils.

Addressing the "Paradox of the Plankton": Using Metabarcoding to Explore Zooplankton Diversity Patterns Across Chemical Conditions in the Salish Sea

Traditional taxonomic analysis of zooplankton is time consuming, expensive, and unable to resolve the true species diversity of a community due to a lack of diagnostic morphological characters for many taxa. This is especially true for early life stages, undescribed, and cryptic species. This limitation has led to a dramatic under-estimation of the incredible diversity of life that inhabits the ocean, hindering our understanding of the environmental conditions that structure communities. Multi-gene metabarcode high-throughput sequencing (HTS) analyses entailing field sampling and bioinformatics offer new means to rapidly and accurately characterize the species identities, diversity, and composition of entire communities. We use multiple diagnostic Illumina MiSeq HTS metabarcode assays (for mitochondrial COI, 16SRNA, and 12SRNA gene regions) and a custom bioinformatics pipeline to analyze communities of invertebrates and fishes from zooplankton net tows collected concurrently with environmental chemistry data across Puget Sound in the southern Salish Sea. We compare results among sites for spring and autumn seasons, as a prelude to multi-year analyses. Findings show considerable divergence in species composition and diversity among sites and season, reflecting differences in salinity, pH, and proximity to the ocean. This approach has great potential for wide-spread use in monitoring programs to assess the diversity of marine plankton communities in conjunction with changing conditions, including ocean acidification, hypoxia, and global temperature rise.

819 Cervicovaginal microbiome and cytokine expression in twin versus singleton gestations

Second trimester cervicovaginal microbiome diversity and cytokine profiles have not been elucidated for twin gestations. We sought to explore differences in second trimester cervicovaginal microbiome diversity and cytokine profiles among women with twin versus singleton gestations. In this prospective cohort study, women with twin gestations were matched to singleton controls without a history of short cervix or preterm birth by gestational age ± 2 weeks and race. Cervicovaginal lavage samples were collected from 14 to 24 weeks gestation during prenatal visits with cervical length if indicated. Cervicovaginal microbiota were analyzed with 16S RNA gene sequencing and classified into community state types (CST) based on Lactobacillus species predominance. A broad survey cytokine multiplexed ELISA panel was performed to assess changes in immune responses. Cytokine profiles and microbiome diversity were compared between groups. 19 twins and 19 singletons had second trimester cervicovaginal microbiome and cytokine analysis performed. Gestational age at sample collection was similar among twins and singletons (19.7 ± 3.4 vs. 19.4 ± 2.9, p=.72). The groups were similar in maternal characteristics, including age, parity, body mass index, preterm birth history and comorbidities. 79% of twin gestations were dichorionic/diamniotic and 21% monochorionic/diamniotic. 3 twin pregnancies were complicated by short cervix compared with 1 singleton pregnancy (p=.6). The cytokine profiles of twins and singletons were not different. Of the 38 cytokines measured, only G-CSF was noted to be elevated in singleton gestations (46.3 [28.2-189.2] vs. 218.5 [47.3-1151] pg/mL, p=.054). CST I, III and V were more prevalent among twin gestations. CST II and IV were similar among the two groups. CST IV was the predominant type among microbiota profiles of twins (55%) and singletons (64%). Second trimester cervicovaginal cytokine profiles were similar among twins and singletons. CST types I, III and V were more prevalent among twins, while rates of CST II and IV were similar to singleton controls.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Heavy-metal resistance mechanisms developed by bacteria from Lerma-Chapala basin.

Heavy-metal (HM) contamination is a huge environmental problem in many countries including Mexico. Currently, microorganisms with multiple heavy-metal resistance and/or plant-promoting characteristics have been widely used for bioremediation of HM-contaminated soils. The aim of the study was isolated bacteria with multiple heavy-metal resistance and to determinate the resistance mechanism developed by these organisms. A total of 138 aerobic bacteria were isolated from soil and sediments surrounding the Lerma-Chapala basin located in the boundary of the States of Michoacán and Jalisco states of Mexico. One hundred and eight strains showed at least 1 plant growth-promoting features. The Lerma-Chapala basin bacteria were also resistant to high concentrations of HMs including the metalloid arsenic. Sequence analysis of 16S RNA genes reveled that these bacteria were mainly affiliated to the phyla Proteobacteria (38%), Firmicutes (31%) and Actinobacteria (25%), covering 21 genera with Bacillus as the most abundant one. Among them, at least 27 putative novel species were detected in the genera Acinetobacter, Arthrobacter, Bacillus, Agrobacterium, Dyadobacter, Enterobacter, Exiguobacterium, Kluyvera, Micrococcus, Microbacterium and Psychrobacter. In addition, these bacteria developed various heavy-metal-resistance mechanisms, such as biosorption/bioaccumulation, immobilization and detoxification. Therefore, the bacteria isolated from soils and sediments of Lerma-Chapala basin could be used in bioremediation strategies.

Rapid detection of fraudulence in seven commercial shrimp products by species-specific PCR assays

Species-specific PCR assay was developed for the authentication of commercially important seven shrimp species. Species-specific primers were designed by targeting the mitochondrial 16S RNA and Internal Transcriber Spacer (ITS1) genes. The PCR assays generated the amplicons of 276, 204, 362, 778, 212, 318 and 330 bp length for Penaeus vannamei, Fenneopenaeus indicus, Penaeus monodon, Macrobrachium rosenbergii, Metapenaeus affinis, Heterocarpus gibbosus,and Penaeus semisulcatus, respectively. The specificity of primers was confirmed with targeted shrimps and non-targeted fishes. The limit of detection of shrimp species was achieved with 1–0.1 ng/μl. The developed assays were validated with 68 commercial shrimp products, and the results revealed 16% of mislabeling. Since, the developed PCR assays possess specificity, sensitivity, and applicability, it can be used as an official method for authentication in shrimp processing industries by the regulatory authorities.

No interplay between gut microbiota composition and the lipopolysaccharide-induced innate immune response in humans in vivo.

ObjectiveAnimal studies have demonstrated the extensive interplay between the gut microbiota and immunity. Moreover, in critically ill patients, who almost invariably suffer from a pronounced immune response, a shift in gut microbiota composition is associated with infectious complications and mortality. We examined the relationship between interindividual differences in gut microbiota composition and variation in the in vivo cytokine response induced by bacterial lipopolysaccharide (LPS). Furthermore, we evaluated whether an LPS challenge alters the composition of the gut microbiota.MethodsHealthy male volunteers received an intravenous bolus of 2 ng kg−1 LPS (n = 70) or placebo (n = 8). Serial plasma concentrations of tumor necrosis factor‐α, interleukin (IL)‐6, IL‐8 and IL‐10 were measured, and subjects were divided into high and low cytokine responders. Gut microbiota composition was determined using 16s RNA gene sequencing of faecal samples obtained 1 day before (baseline) and 1 day and 7 days following the LPS challenge.ResultsBaseline microbiota composition, analysed by principal coordinate analysis and random forest analysis, did not differ between high and low responders for any of the four measured cytokines. Furthermore, baseline microbiota diversity (Shannon and Chao indices) was similar in high and low responders. No changes in microbiota composition or diversity were observed at 1 and 7 days following the LPS challenge.ConclusionOur results indicate that existing variation in gut microbiota composition does not explain the observed variability in the LPS‐induced innate immune response. These findings strongly argue against the interplay between the gut microbiota composition and the innate immune response in humans.

Gut Microbiota Composition in CRC Patients with Liver Metastasis

Introduction: Liver metastases contribute to the high mortality rates of colorectal cancer (CRC).Until now, no study has reported the gut microbiota composition in CRC patients with liver metastasis. Method: We conducted a cross-sectional and case-control study, to analysis the gut microbiota of CRC with synchronous liver metastasis patients at initial diagnosis (sLM) and patients who develop metachronous liver metastasis (mLM) after surgery, compared to patients without liver metastasis (CG). Gut microbiota were analyzed by 16s RNA gene sequencing with mucosal samples. Result: An analysis of the mucosal microbial diversity with two methodologies (Shannon and Simpson indices) showed that alpha-diversity of the mucosal microbiome was significantly higher in CG group compared with that of sLM or mLM groups.Principal coordinate analysis (PCoA) on geuns level with Bary-Curtis distance showed a separated gut microbiota of both sLM and mLM from CG. LEfSe analysis identified 5 predominated taxa between the sLM and CG groups, 12 predominated taxa between the mLM and CG groups on genus level with LAD score>3.0, It was noted that Escherichia_Shigella had higher relative abundance in the Both sLM and mLM groups compared to the CG group (P< 0.05) Conclusion: CRC patients with synchronous or metachronous liver metastasis showed a different gut microbiota. Whether gut microbiota contribute to liver metastasis need further validation.

Identification of No. 8 spoilage bacteria and study on biological properties of areca taro

You should leave 8 mm of space above the abstract and 10 mm after the abstract. The heading Abstract should be typed in bold 9-point Times. The body of the abstract should be typed in normal 9-point Arial in a single paragraph, immediately following the heading. The text should be set to 1 line spacing. The abstract should be centred across the page, indented 17 mm from the left and right page margins and justified. It should not normally exceed 200 words. In order to explore the effects of fungus infestation on areca taro during cold storage period, a black morphologic fungus (No. 8) was isolated and identified as Curvularia lunata by morphological observation and 18sRNA gene sequencing. The study on its biological properties showed that the growth and development temperature of Curvularia lunata were 15-40 ℃, the optimal temperature was 25-30 ℃, the optimal pH was 6-8, the optimal light condition was natural light, the optimal carbon source was sorbitol, maltodextrin and mannitol, and the optimal nitrogen source was glycine. The optimum germination temperature, pH and humidity of conidia were 25 - 40℃, 6 - 9 and 100% + water drop, respectively, under the promotion of 40% areca taro solution. The aim of this study was to provide a theoretical basis for the prevention and treatment of areca taro rotting in cold storage period.

716. A Case of Symptomatic Intestinal Spirochetosis

BackgroundIntestinal spirochetosis (IS) is a condition caused by Brachyspira aalborgi and Brachyspira Pilosicoli. Its clinical significance has long been a point of contention with some debating that these spirochetes are simply colonic commensals. It is a condition that is more prevalent in developing nations as well as patients with HIV and the homosexual population. The epidemiology and prevalence of IS has not been studied in the local context.MethodsWe reviewed a case of a 37-year-old man who presented with a two month history of persistent lower abdominal pain, hematochezia, and increase in mucous discharge per rectum. He is sexually active with multiple male partners, and was previously treated for gonorrhea, chlamydia, and syphilis. His basic laboratory investigations were unremarkable, Venereal Disease Research Laboratory (VDRL) antibody and human immunodeficiency virus (HIV) screen were both non-reactive. Computed tomography of the abdomen was unremarkable. Endoscopic evaluation revealed multiple discrete ulcers measuring 1-2mm seen only in the rectum. Random biopsies of the cecum, ascending colon, transverse colon and descending colon showed mild acute colitis with IS. There was also mild to moderate acute proctitis in the rectum with spirochetes seen. 16s RNA gene sequencing of the biopsy specimen were confirmatory for Brachyspira aalborgi.Investigation findings. A: Discrete Ulcers found in rectum, B: Hemotoxylin and Eosin stained specimen showing proctitis, C: False brush Border appearance D: Spirochetes on Warthin Starry stain ResultsThe patient received a 10 day course of metronidazole with complete resolution of his symptoms.ConclusionThis case demonstrates the existence of a treatable condition that can be diagnosed with current available investigations for patients with similar symptoms. Recognising at risk populations can also raise clinical suspicion for this condition. Some studies have found associations between IS with development of colonic polyps and also certain colorectal cancers. Further studies on this treatable condition and its disease burden in the local context should be further explored.Disclosures All Authors: No reported disclosures

Isolation and Molecular Identification of Nontuberculous Mycobacterium from Different Species of Fish in Karbala Province, Iraq

This study was designed for isolation and molecular identification of Nontuberculous Mycobacterium (NTM) from fish during the period between October and December 2017 from Karbla province, Iraq. This study included 200 fresh fish samples from four different species including Spondyliosoma cantharus, Liza abu, Carassius carassius and Cyprinuscarpio. Three samples of each fish were taken including gills, muscles and all internal organs. The samples were processed by decontamination, concentration of 4% sodium hydroxide, and 0.1 ml of sediment was streaking on Löwenstein Johnson (LJ) media; then the bacterial cultures were incubated at 28-30 °C for 3days up to 4 weeks and suspected colonies were stained with acid fast stain to confirm the presence of Mycobacterium. Further identification, biochemical tests were carried out to confirm the diagnosis of isolates, PCR was done using 16s RNA gene for all isolates, hsp65 gene was used in unidentified NTM spp and to confirm the others. Results revealed that out of 200 fish samples, 19 isolates 9.5% were identified as NTM belonged to Rapid Growth Mycobacterium (RGM). of the total isolates, 18.26 % was investigated from Liza abu (Kishni, Abu khraiza). NTM (RGM) isolates on spp level identified six spp of these isolates. M. porcinum was 26.32% which was followed by M. fortuitum of 21.05%, others included M. neworleansense and M. mucogenicum 10.5% of each, M. cosmeticum and M. pallens 5.26% of each. The distribution of NTM spp in the fish organs, nine out of 19 (47.37%) NTM isolate were recovered from gills followed by muscles 36.84 %, while 15.79% from internal organs. These results were the first study concerning isolation of these spp of NTM from fish in Iraq, and some spp are not reported in other studies. This study concluded that the fish is an importance source or reservoir for NTM, especially the pathogenic spp.

Disparate outer membrane exclusionary properties underlie intrinsic resistance to hydrophobic substances in Pseudomonas spp. isolated from surface waters under triclosan selection

Representative members of surface water microbiota were obtained from three unrelated municipal sites in Oklahoma by direct plating under selection by the hydrophobic biocide triclosan. Multiple methods were employed to determine if intrinsic triclosan resistance reflected resistance to hydrophobic molecules by virtue of outer membrane impermeability. While all but one organism isolated in the absence of triclosan were able to initiate growth on MacConkey agar, only one was able to initiate significant growth with triclosan present. In contrast, all bacteria selected with triclosan were identified as Pseudomonas spp. using 16S RNA gene sequencing and exhibited growth comparable to Pseudomonas aeruginosa controls in the presence of hydrophobic antibacterial agents to include triclosan. Two representative bacteria isolated in the absence of triclosan allowed for greater outer membrane association with the fluorescent hydrophobic probe 1-N-phenylnapthylamine than did two triclosan-resistant isolates. Compound 48/80 disruption of outer membrane impermeability properties for hydrophobic substances either partially or fully sensitized nine of twelve intrinsically resistant isolates to triclosan. These data suggest that outer membrane exclusion underlies intrinsic resistance to triclosan in some, but not all Pseudomonas spp. isolated by selection from municipal surface waters and implicates the involvement of concomitant triclosan resistance mechanisms.

Correlations between microbiota and metabolites after faecal microbiota transfer in irritable bowel syndrome

Faecal microbiota transfer (FMT) consists of the infusion of donor faecal material into the intestine of a patient with the aim to restore a disturbed gut microbiota. In this study, it was investigated whether FMT has an effect on faecal microbial composition, its functional capacity, faecal metabolite profiles and their interactions in 16 irritable bowel syndrome (IBS) patients. Faecal samples from eight different time points before and until six months after allogenic FMT (faecal material from a healthy donor) as well as autologous FMT (own faecal material) were analysed by 16S RNA gene amplicon sequencing and gas chromatography coupled to mass spectrometry (GS-MS). The results showed that the allogenic FMT resulted in alterations in the microbial composition that were detectable up to six months, whereas after autologous FMT this was not the case. Similar results were found for the functional profiles, which were predicted from the phylogenetic sequencing data. While both allogenic FMT as well as autologous FMT did not have an effect on the faecal metabolites measured in this study, correlations between the microbial composition and the metabolites showed that the microbe-metabolite interactions seemed to be disrupted after allogenic FMT compared to autologous FMT. This shows that FMT can lead to altered interactions between the gut microbiota and its metabolites in IBS patients. Further research should investigate if and how this affects efficacy of FMT treatments.

Metagenomic Approach to Bacterial Diversity and Lipolytic Enzymes’ Genes from a Steam Soil of Los Humeros Geothermal Field (Puebla, México)

Los Humeros volcanic complex, located at the eastern end of the Trans Mexican Volcanic Belt, possess superficial thermal manifestations (fumaroles, heat steam-soils and alteration zones), with temperatures between 50 °C and 90 °C. This is the first work to explore the bacterial diversity of a steam soil of Los Humeros geothermal field using a metagenomic 16S RNA gene analysis, and the presence of lipolytic enzymes’ genes after an enrichment culture by a whole-genome shotgun approach. High diversity indexes were found in soil samples, with Firmicutes, Chloroflexi, Proteobacteria, Actinobacteria and Acidobacteria as the most prevalent phyla. Alicyclobacillus was the most prevalent genus at 15 cm and Anoxybacillus at 50 cm depth; anaerobic and obligate thermophilic and hyperthermophilic members were found at 50 cm depth. Firmicutes was favored by the oil-rich medium, and the dominant genera were Geobacillus and Caldibacillus. 165 genes related to lipolytic enzymes were found; carboxilesterases and phospholipases C were the most abundant, and 43% were assigned to uncharacterized lipolytic enzymes. The diversity as well as the high percentage of non-characterized lipolytic enzymes found after soil enrichment in an oil-rich medium suggest the presence of biotechnologically promising organisms and enzymes that deserve to be investigated.

Gut Microbiota of Young Children Living in Four Brazilian Cities.

Recent studies have demonstrated that gut microbiota development is influenced by human biogeographic factors such as race, ethnicity, diet, lifestyle or culture-specific variations, and other environmental influences. However, biogeographic variation in gut microbiota assembly remains largely unexplored in Latin America. In this paper, we compared food recall information and microbiota composition of toddlers living in geographically separated urban populations within four states of Brazil. 16S RNA gene sequencing revealed that alpha diversity was similar between the four different populations. Gut microbiota compositions were dominated by members of the phyla Firmicutes and Bacteroidetes, resembling a more adult-like microbiota as compared with those of Western European toddlers of similar age. These findings suggest that inter-individual and nutrition-induced differences were apparent in the fecal microbiota. We conclude that urban dietary pattern plays a larger role in influencing the gut microbiota composition than do biogeographic factors.

Total nitrogen removal in biochar amended non-aerated vertical flow constructed wetlands for secondary wastewater effluent with low C/N ratio: Microbial community structure and dissolved organic carbon release conditions

Biochar was utilized to intensify constructed wetland (CW) for further organic and nitrogen removal from secondary wastewater. Four sets of non-aerated biochar amended vertical flow CW (VFCW) were developed to investigate the synergistic effects of biochar and microbes on pollutant removal. Results showed that the average COD and nitrogen removal efficiencies of VFCW1 (with 1% w/w biochar with microbe and plants) achieved 89.1 ± 5.6% and 90.2 ± 3.1% respectively, and their corresponding removal rates of 10.2 ± 0.8 mg-COD/(m3.d) and 3.57 ± 0.3 mg-TN/(m3.d) which were 35 and 52.3% higher than control. The biochar’s dissolved organic carbon release in VFCWs indicated that water and acidic media portray the optimum conditions for nitrogen removal. The 16S RNA gene sequencing analysis indicated that in the biochar-amended VFCWs, bacterial phylum Proteobacteria (24.13–51.95%) followed by Chloroflexi (5.64–25.01%), Planctomycetes (8.48–14.43%), Acidobacteria (2.29–11.65%) were abundantly enhanced. Conclusively, incorporating biochar in non-aerated VFCWs is an efficient technique for enhancing nitrogen removal from secondary effluent.

Effects of Ocean Acidification and Microplastics on Microflora Community Composition in the Digestive Tract of the Thick Shell Mussel Mytilus coruscusThrough 16S RNA Gene Sequencing.

Ocean acidification and microplastic pollution is a global environmental threat, this research evaluated the effects of ocean acidification and microplastics on mussel digestive tract microbial community. The 16S rRNA gene was sequenced to characterize the flora. Species diversity in the samples was assessed by clustering valid tags on 97% similarity. Bacteroidetes, Firmicutes and Proteobacteria were the three most abundant genera in the four groups, with Bacteroidetes showing the highest diversity. However, no differences in flora structure were evident under various treatments. Phylogenetic relationship analysis revealed Bacteroidetes and Firmicutes had the highest OTU diversity. The weighted UniFrac distance, principal coordinate analysis (PCoA), unweighted pair group method with arithmetic mean (UPGMA) cluster tree and analysis of molecular variance (AMOVA) evaluation results for all samples also showed that changes in pH and microplastics concentration did not significantly affect the microbial community structure in the mussel digestive tract. The results presented the no significant effects of ocean acidification and microplastics intake on mussel intestinal diversity.

MALDI-TOF MS and 16S RNA Identification of Culturable Gastric Microbiota: Variability Associated with the Presence of Helicobacter pylori.

Helicobacter pylori is the main bacteria associated with gastroduodenal diseases. Recent studies have reported that gastric microbiota might be modified by the H. pylori colonization, favoring gastric lesions′ development. In Chile, the region of La Araucanía concentrates a high risk of gastric cancer associated with Helicobacter pylori colonization, rurality, poverty, and Mapuche ethnicity. Hence, we aimed to identify the culturable gastric microbiota and characterize its variability at different stages of epithelial injury, based on its H. pylori colonization in dyspeptic patients from this Chilean region. Microaerophilic bacteria strains were isolated from antrum biopsies of 155 dyspeptic patients′ biopsies and identified using MALDI-TOF MS or 16sRNA gene sequencing for non-pylori species identification, and UreC gene amplification for H. pylori confirmation. We found 48 species from 18 families, mainly belonging to Neisseriaceae (21.3%), Streptococcaceae (20.0%), Actynomicetaceae (9.0%), Enterobacteriaceae, and Lactobacillaceae (4.5%); however, Streptococcaceae and Actinomycetaceae families showed a significant reduction in samples infected with H. pylori, along with a considerably lower diversity of species. Our results revealed a microbiota modification due to H. pylori colonization associated with the gastric epithelial state, suggesting a potential microbiota role for developing and progressing gastric diseases.

Gut Bacterial Diversity Associates with Efficacy of Anti-CD19 CAR T-Cell Therapy in Patients with Large B-Cell Lymphoma

Gut Bacterial Diversity Associates with Efficacy of Anti-CD19 CAR T-Cell Therapy in Patients with Large B-Cell Lymphoma

Coptis chinensis Franch polysaccharides provide a dynamically regulation on intestinal microenvironment, based on the intestinal flora and mucosal immunity

Ethnopharmacological relevanceCoptis chinensis Franch is one of the most widely used traditional Chinese herbs in China and was firstly recorded in "Shennong's Classic of Materia Medica” in the Han Dynasty. The medical records in past thousands years have fully confirmed the clinical efficacies of Coptis chinensis Franch against intestinal diseases. The polysaccharides in herbal medicines can be digested by the flora and uptaken by the Peyer's patches (PPs) in intestine. It can be reasonably presumed that the polysaccharides in Coptis chinensis Franch (CCP) should be one of the critical element in the regulation of intestinal microenvironment. Aim of the studyThis study intended to explore the dynamic regulation of CCP on intestinal microenvironment from the perspective of the intestinal mucosal immunity and the intestinal flora, in order to provide a new research perspective for the pharmacological mechanism of Coptis chinensis Franch. Materials and methodsThe absorption and distribution of CCP in intestinal tissues were observed after the perfusion of FITC labeled CCP. The influences of CCP on intestinal flora were evaluated by the 16sRNA gene illumina-miseq sequencing after gavage. The regulations of CCP on intestinal mucosal immunity were evaluated by the immunohistochemical analysis of the interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-17 (IL-17) and transforming growth factor-β (TGF-β) secretion in PPs and intestinal epithelial tissue. ResultsWith the self-aggregation into particles morphology, CCP can be up-taken by PPs and promote the IFN-γ, IL-4, IL-17 and TGF-β secretion in PPs in a dose-dependent manner. The CCP can also be utilized by the intestinal flora and dynamically regulate the diversity, composition and distribution of the intestinal flora. The temporal regulations of CCP on IFN-γ, IL-4, IL-17 and TGF-β secretions in intestinal epithelial tissues are consistent with the variation tendency of intestinal flora. ConclusionCCP can provide effective, dynamical and dose-dependent regulations on intestinal microenvironment, not only the intestinal flora but also the PPs and intestinal epithelium related immune response. These may be involved in the multiple biological activities of Coptis chinensis Franch.